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5-氮杂-2’-脱氧胞苷和4-苯基丁酸对慢性粒细胞白血病细胞的miR-196b表达水平有协同作用
刘玥 帅春 李杰生 尹虹 宋艳斌 马文丽*
解剖学报 ›› 2014, Vol. 45 ›› Issue (4) : 521-524.
5-氮杂-2’-脱氧胞苷和4-苯基丁酸对慢性粒细胞白血病细胞的miR-196b表达水平有协同作用
5-Aza-2’-deoxycytidine and 4-phenylbutyric acid exert a cooperative effect on the expression of miR-196b in chronic myeloid leukemia cells
目的 探讨慢性粒细胞白血病细胞中影响miR-196b表达水平的表观遗传学因素。方法 分别采用DNA甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷（5-Aza-2’-dc）、组蛋白去乙酰化酶抑制剂4-苯基丁酸（PBA）和两者联合处理K562细胞，采用实时定量PCR（Real-time PCR）检测miR-196b的表达水平变化。
结果 PBA的半数抑制浓度为1.58mmol/L。和正常人骨髓细胞miR-196b的表达水平相比，Aza组、PBA组和阴性对照组miR-196b的表达水平显著降低且基本一致，Aza＋PBA组miR-196b的表达水平和正常人表达水平基本一致。结论 单独使用 5-Aza-2’-dc或PBA不能使K562细胞中miR-196b的表达恢复正常，两者联合使用共同处理K562细胞，可使miR-196b的表达恢复正常，表明K562细胞中miR-196b的表达水平和基因组甲基化及组蛋白乙酰化均有关系。
Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells.
Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR. Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L. Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group, PBA group and negative control cells and nearly consistent among three groups, and as high as normal cells in combined treatment group. Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2’-deoxycytidine or 4-phenylbutyric acid separately, while could restore normal when treated with both agents, indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation.
miR-196b / 甲基化 / 组蛋白乙酰化 / K562细胞 / 实时定量聚合酶链反应 / 人
miR-196b / Methylation / Histone acetylation / K562cell / Real-time PCR / Human
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广东省医学科研基金;南方医科大学2012年度“科研启动计划”青年科技人员培育项目;广东省科技计划项目;广东省自然基金项目
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